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Clinical and Diagnostic Laboratory Immunology, July 2002, p. 883-891, Vol. 9, No. 4
1071-412X/02/$04.00+0     DOI: 10.1128/CDLI.9.4.883-891.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Use of Multiepitope Polyproteins in Serodiagnosis of Active Tuberculosis

Raymond L. Houghton,1* Michael J. Lodes,1 Davin C. Dillon,1 Lisa D. Reynolds,1 Craig H. Day,1 Patricia D. McNeill,1 Ronald C. Hendrickson,1 Yasir A. W. Skeiky,1 Diana P. Sampaio,2,3 Roberto Badaro,2,3 Konstantin P. Lyashchenko,4 and Steven G. Reed1

Corixa Corporation, Seattle, Washington,1 Chembio Diagnostics Systems, Inc., Medford, New York,4 Federal University of Bahia,2 Fundacao Bahiana de Infectologia, Salvador, Brazil3

Received 17 January 2002/ Returned for modification 25 March 2002/ Accepted 11 April 2002

Screening of genomic expression libraries from Mycobacterium tuberculosis with sera from tuberculosis (TB) patients or rabbit antiserum to M. tuberculosis led to the identification of novel antigens capable of detecting specific antibodies to M. tuberculosis. Three antigens, Mtb11 (also known as CFP-10), Mtb8, and Mtb48, were tested together with the previously reported 38-kDa protein, in an enzyme-linked immunosorbent assay (ELISA) to detect antibodies in TB patients. These four proteins were also produced as a genetically fused polyprotein, which was tested with two additional antigens, DPEP (also known as MPT32) and Mtb81. Sera from individuals with pulmonary and extrapulmonary TB, human immunodeficiency virus (HIV)-TB coinfections, and purified protein derivative (PPD)-positive and PPD-negative status with no evidence of disease were tested. In samples from HIV-negative individuals, the ELISA detected antibodies in >80% of smear-positive individuals and >60% smear-negative individuals, with a specificity of ~98%. For this group, smears detected 81.6% but a combination of smear and ELISA had a sensitivity of ~93%. The antigen combination detected a significant number of HIV-TB coinfections as well as antibodies in patients with extrapulmonary infections. Improved reactivity in the HIV-TB group was observed by including the antigen Mtb81 that was identified by proteomics. The data indicate that the use of multiple antigens, some of which are in a single polyprotein, can be used to facilitate the development of a highly sensitive test for M. tuberculosis antibody detection.

* Corresponding author. Mailing address: Corixa Corporation, 1124 Columbia St., Suite 200, Seattle, WA 98104. Phone: (206) 754-5747. Fax: (206) 754-5917. E-mail: houghton{at}corixa.com.

Clinical and Diagnostic Laboratory Immunology, July 2002, p. 883-891, Vol. 9, No. 4
1071-412X/02/$04.00+0     DOI: 10.1128/CDLI.9.4.883-891.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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